Isolation, Purification and Identification of Indigenous Bacteria from PAHs Polluted Soil

Document Type : Research Paper



PAHs are toxic compounds with carcinogenic effects on humans that are released into the environment by incomplete combustion of fossil fuels. Three methods are commonly employed for PAHs pollutant removal: physical, chemical, and biological. From among these, the biological method which typically contains microbial processes and transforms pollutants to nontoxic or less toxic substances is the most innocuous and effective solution. In this study, attempts were initially made to enrich, isolate, and purify indigenous bacteria from PAHs polluted soil. In the second stage, the PCR method was exploited to identify the bacteria that had the capability of growth and reproduction in polluted conditions. It was found that the degrading bacteria are component species of gram negative bacilli determined as Shewanella, Pseudomonas, and Achromobacter. The results of the present study indicate that the bacteria have the best performance in PAHs removal from polluted environments.


Main Subjects

1. Nie, M., Xiao, Z., Jin, W., and Chang, F. (2009). “Rhizosphere effects on soil bacterial abundance and diversity in the Yellow River Deltaic ecosystem as influenced by petroleum contamination and soil salinization.” J. Soil Biology and Biochemistry, 41, 2535-2542.
2. Carsten Suhr, J. (1997). “Plant protection and rhizosphere colonization of barley by seed inoculated herbicide degrading Burkholderia (Pseudomonas) in 2,4-D contaminated soil.” J. Plant and Soil, 189, 139-144.
3. Kaksonen1, A.H., Jussila, M.M., Lindstro¨m, K., and Suominen, L.  (2006). “Rhizosphere effect of Galega Orientalis in oil-contaminated soil.” Soil Biology and Biochemistry, 38, 817-827.
4. Johnsona, D.L., Andersonb, D.R., and McGratha, S.P. (2005). “Soil microbial response during the phytoremediation of a PAH contaminated soil.” J. Soil Biology and Biochemistry, 37, 2334-2336.
5. Baek, S.O., Field, R.A., Goldstone M.E., Kirk P.W., and Lester, J.N. (1991). “A review of atmospheric poly cyclic aromatic  hydocarbonse: Sources , fate and behavior.” Water Air Soil pollut., 60, 279-300.
6. Niel, O.P. (1995). Environment chemistry, 2nd Ed., Chapman and Hall London SE1 8HN, UK, p237.
7. World Health Organization. (1998). Selected non heterocyclic poly cyclic aromatic hydrocarbons environmental health criteria 202 International program on chemical safty WHO, Geneva.
8. US.EPA. (1997). Code of federal regulation title 40 part 60 subparts D Da Db Dc, Environmental Protection Agency Washington DC. 44.
9. Yu, H., and Xue, Y. Y. (2009). “Interactions between selected PAHs and the microbial community in rhizosphere of a paddy soil.” Science the Total Environment, 407, 1027-1034.
10. Haritash, A.K., and Kaushid, C.P. (2009). “Biodegradation aspects of polyaromatic hydrocarbons (PAHs).” J. of Hazardous Materials, 169, 1-15.
11. Aitken, M.D., Stringfellow, W.T., Nagel, R.D., Kazunga, C., and Chen, S.H. (1998). “Characteristics of phenanthrene-degrading bacteria isolated from soils contaminated with polycyclic aromatic hydrocarbons.” Canadian Journal of Microbiology, 44, 743-752.
12. Rhoades, J. D. (1986). “Cation exchange capacity.” Page, A. C. (Ed). Methods of soil analysis, part. 2, American Society Agronomy, 9.
13. Rodrigo, S., Okeke, C., Peralba, M., and Camargo, O. (2009). “Smproved enrichment and iohtion of polycyclic aromatic hydrocerbos PAH-degrading microorgomisn in soil using Anhracene as arnodel PAH. Curr. Microbiology, 58, 628-634.
14. Sang-Hwan Lee, A., Won-Seok Lee, B., Chang-Ho Lee, C., and Jeong-Gyu, K. (2008). “Degradation of phenanthrene and pyrene in rhizosphere of grasses and legumes.” J. of Hazardous Materials, 153 892-898.
15. Neuman, G., Teras, R., Monson, L., Kivisaar, M., Schauer, F., and Heipieper, J. (2004). “Simultaneous degradation of atrazine and phenol by Pseudomonas sp. Strain ADP: Effects of toxicity and adaptation.” Applied and Environmental Microbiology Journal, 70, 1907-1912.
16. Eskandary, S., Tahmourespour, A., and Hoodaji, M. (2011). “Investigation of growth and removal of phenol by isolation of bacterium from industrial waste water in vitro.” J. Water and Wastewater, 78, 78-85. (In Persian)
17. Holt, G., Krieg R., Sneath, A., Staley, T., and Williams, T. (1994). Bergeys manual of d eterminative bacteriology, 9 th Ed., Williams and Wilkins Publisher, Baltimor.
18. Mcpherson, M.J., and Muller, S.G. (2000). PCR, Bios Scientific Publisher, New York, Oxford.
19. United State Environmenal Protection Agency. (2008). Poly cyclic aromatic hydrocarbonse, USEPA.
20. Johnsona, D.L., Andersonb, D.R., and McGratha, S.P. (2005). “Soil microbial response during the phytoremediation of a PAH contaminated soil.” J. Soil Biology and Biochemistry, 37, 2334-2336.
21.Ye, D., Akmail Siddiqi, M., Maccubbin, A., Kumar, S., and Sikra, H. (1996). “Degradation of polynuclear aromatic hydrocarbons by sphingomonas paucimobilis.” Environmental Science and Technology, 30,
22. Romero, M.C., Cazau, M.C., Giorgieri, S., and Arambarri, A.M. (1998). “Phenanthrene degradation by microorganisms isolated from a contaminated stream.” Environmental Pollution, 101, 355-359.
23. Yuan, S.Y., Chang, S.W., and Chang B.V. (2003) “Biodegradation of polycyclic aromatic hydrocarbons in sludge. Bulletin of Environmental Contamination and Toxicology, 71, 625-632.
24. Dean-Ross.” D., Moody, J.D., Freeman, J.P., Doerge, D.R., and Cerniglia, C.E. (2001). “Metabolism of anthracene by a rhodococcus species FEMS.” Microbiology Letters, 204, 205-211.
25. Gentile, G., Bonasera, V., Amico, C., Giuliano, L., and Yakimov, M.M. (2003) “Shewanella sp. GA-22, a psychrophilic hydrocarbonoclastic antarctic bacterium producing polyunsaturated fatty acids.” J. Appl. Microbiol, 95, 1124-1133.
26. Roda, F., Desouky, A. M., and Al-Shammri, M. (2009). “Isolation and characterization of polyaromatic hydrocarbons-degrading bacteria from different Qatari soils.” African Journal of Microbiology Research,3(11), 761-766.